Monitoring of the prostate tumour cells redox state and
real-time proliferation by novel biophysical techniques and
fluorescent staining

J 2012

Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining

MASAŘÍK, Michal, Jaromír GUMULEC, Marián HLAVNA, Markéta SZTALMACHOVÁ, Petr BABULA et. al.

Basic information

Original name

Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining

Authors

MASAŘÍK, Michal (203 Czech Republic, guarantor, belonging to the institution), Jaromír GUMULEC (203 Czech Republic, belonging to the institution), Marián HLAVNA (703 Slovakia, belonging to the institution), Markéta SZTALMACHOVÁ (203 Czech Republic, belonging to the institution), Petr BABULA (203 Czech Republic), Martina RAUDENSKÁ (203 Czech Republic, belonging to the institution), Monika PÁVKOVÁ GOLDBERGOVÁ (203 Czech Republic, belonging to the institution), Natalia Vladimirovna CERNEI (498 Republic of Moldova), Jiří SOCHOR (203 Czech Republic), Ondřej ZÍTKA (203 Czech Republic), Branislav RUTTKAY-NEDECKÝ (203 Czech Republic), Soňa KŘÍŽKOVÁ (203 Czech Republic), Vojtěch ADAM (203 Czech Republic) and René KIZEK (203 Czech Republic)

Edition

Integrative Biology, 2012, 1757-9694

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30000 3. Medical and Health Sciences

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 4.321

RIV identification code

RIV/00216224:14110/12:00057446

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.1039/c2ib00157h

UT WoS

000304487300010

Keywords in English

OXIDATIVE STRESS; CANCER CELLS; LIQUID-CHROMATOGRAPHY; CARCINOMA-CELLS; ACRIDINE-ORANGE; ZINC; METALLOTHIONEIN; DIFFERENTIATION; TRANSPORTER; GLUTATHIONE

Abstract

V originále

The present paper is focused on zinc(II) treatment effects on prostatic cell lines PC-3 (tumour) and PNT1A (non-tumour). Oxidative status of cells was monitored by evaluation of expression of metallothionein (MT) isoforms 1A and 2A at the mRNA and protein level, glutathione (oxidised and reduced), and intracellular zinc(II) after exposition to zinc(II) treatment at concentrations of 0–150 mM using electrochemical methods, western blotting and fluorescent microscopy. A novel real-time impedance-based growth monitoring system was compared with widely used end-point MTT assay. Impedance-based IC50 for zinc(II) is 55.5 and 150.8 mM for PC-3 and PNT1A, respectively. MTTdetermined IC50 are >1.3-fold higher. Impedance-based viability correlates with viable count (r > 0.92; p o 0.03), not with MTT. Two-fold lower intracellular zinc(II) in the tumour PC-3 cell line was found. After zinc(II) treatment >2.6-fold increase of intracellular zinc(II) was observed in non-tumour PNT1A and in tumour PC-3 cells. In PC-3 cells, free and bound zinc(II) levels were enhanced more markedly as compared to PNT1A. PNT1A produced 4.2-fold less MT compared to PC3. PNT1A cells showed a 4.8-fold increase trend (r = 0.94; p = 0.005); PC-3 did show a significant trend at MT1 and MT2 protein levels (r = 0.93; p = 0.02) with nearly ten-fold increase after 100 mM zinc(II) treatment. In terms of redox state, PNT1A had a predominance of reduced GSH forms (GSH : GSSG ratio > 1), when exposed to zinc(II) compared to PC3, where predominance of oxidised forms remains at all concentrations. IC50 differs significantly when determined by MTT and real-time impedance-based assays due to dependence of impedance on cell morphology and adhesion. When real-time growth monitoring, precise electrochemical methods and fluorescent microscopy are performed together, accurate information for metal fluxes, their buffering by thiol compounds and monitoring of the redox state become a powerful tool for understanding the role of oxidative stress in carcinogenesis.

Links

GP301/09/P436, research and development project

Name: Analýza metalothioneinu u karcinomu prostaty na úrovni DNA, RNA a proteinu.

Investor: Czech Science Foundation

MUNI/A/0839/2011, interní kód MU

Name: Patofyziologické aspekty vybraných genotypů a fenotypů komplexních nemocí (Acronym: Patofyziologie komplexních nemocí)

Investor: Masaryk University, Category A

Citovat

MASAŘÍK, Michal, Jaromír GUMULEC, Marián HLAVNA, Markéta SZTALMACHOVÁ, Petr BABULA, Martina RAUDENSKÁ, Monika PÁVKOVÁ GOLDBERGOVÁ, Natalia Vladimirovna CERNEI, Jiří SOCHOR, Ondřej ZÍTKA, Branislav RUTTKAY-NEDECKÝ, Soňa KŘÍŽKOVÁ, Vojtěch ADAM and René KIZEK. Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining. Integrative Biology. 2012, vol. 4, No 6, p. 672–684. ISSN 1757-9694. Available from: https://dx.doi.org/10.1039/c2ib00157h.

@article{984961,
   author = {Masařík, Michal and Gumulec, Jaromír and Hlavna, Marián and Sztalmachová, Markéta and Babula, Petr and Raudenská, Martina and Pávková Goldbergová, Monika and Cernei, Natalia Vladimirovna and Sochor, Jiří and Zítka, Ondřej and RuttkayandNedecký, Branislav and Křížková, Soňa and Adam, Vojtěch and Kizek, René},
   article_number = {6},
   doi = {http://dx.doi.org/10.1039/c2ib00157h},
   keywords = {OXIDATIVE STRESS; CANCER CELLS; LIQUID-CHROMATOGRAPHY; CARCINOMA-CELLS; ACRIDINE-ORANGE; ZINC; METALLOTHIONEIN; DIFFERENTIATION; TRANSPORTER; GLUTATHIONE},
   language = {eng},
   issn = {1757-9694},
   journal = {Integrative Biology},
   title = {Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining},
   volume = {4},
   year = {2012}
}

TY  - JOUR
ID  - 984961
AU  - Masařík, Michal - Gumulec, Jaromír - Hlavna, Marián - Sztalmachová, Markéta - Babula, Petr - Raudenská, Martina - Pávková Goldbergová, Monika - Cernei, Natalia Vladimirovna - Sochor, Jiří - Zítka, Ondřej - Ruttkay-Nedecký, Branislav - Křížková, Soňa - Adam, Vojtěch - Kizek, René
PY  - 2012
TI  - Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining
JF  - Integrative Biology
VL  - 4
IS  - 6
SP  - 672–684
EP  - 672–684
SN  - 17579694
KW  - OXIDATIVE STRESS
KW  - CANCER CELLS
KW  - LIQUID-CHROMATOGRAPHY
KW  - CARCINOMA-CELLS
KW  - ACRIDINE-ORANGE
KW  - ZINC
KW  - METALLOTHIONEIN
KW  - DIFFERENTIATION
KW  - TRANSPORTER
KW  - GLUTATHIONE
N2  - The present paper is focused on zinc(II) treatment effects on prostatic cell lines PC-3 (tumour) and PNT1A (non-tumour). Oxidative status of cells was monitored by evaluation of expression of metallothionein (MT) isoforms 1A and 2A at the mRNA and protein level, glutathione (oxidised and reduced), and intracellular zinc(II) after exposition to zinc(II) treatment at concentrations of 0–150 mM using electrochemical methods, western blotting and fluorescent microscopy. A novel real-time impedance-based growth monitoring system was compared with widely used end-point MTT assay. Impedance-based IC50 for zinc(II) is 55.5 and 150.8 mM for PC-3 and PNT1A, respectively. MTTdetermined IC50 are >1.3-fold higher. Impedance-based viability correlates with viable count (r > 0.92; p o 0.03), not with MTT. Two-fold lower intracellular zinc(II) in the tumour PC-3 cell line was found. After zinc(II) treatment >2.6-fold increase of intracellular zinc(II) was observed in non-tumour PNT1A and in tumour PC-3 cells. In PC-3 cells, free and bound zinc(II) levels were enhanced more markedly as compared to PNT1A. PNT1A produced 4.2-fold less MT compared to PC3. PNT1A cells showed a 4.8-fold increase trend (r = 0.94; p = 0.005); PC-3 did show a significant trend at MT1 and MT2 protein levels (r = 0.93; p = 0.02) with nearly ten-fold increase after 100 mM zinc(II) treatment. In terms of redox state, PNT1A had a predominance of reduced GSH forms (GSH : GSSG ratio > 1), when exposed to zinc(II) compared to PC3, where predominance of oxidised forms remains at all concentrations. IC50 differs significantly when determined by MTT and real-time impedance-based assays due to dependence of impedance on cell morphology and adhesion. When real-time growth monitoring, precise electrochemical methods and fluorescent microscopy are performed together, accurate information for metal fluxes, their buffering by thiol compounds and monitoring of the redox state become a powerful tool for understanding the role of oxidative stress in carcinogenesis.
ER  -

MASAŘÍK, Michal, Jaromír GUMULEC, Marián HLAVNA, Markéta SZTALMACHOVÁ, Petr BABULA, Martina RAUDENSKÁ, Monika PÁVKOVÁ GOLDBERGOVÁ, Natalia Vladimirovna CERNEI, Jiří SOCHOR, Ondřej ZÍTKA, Branislav RUTTKAY-NEDECKÝ, Soňa KŘÍŽKOVÁ, Vojtěch ADAM and René KIZEK. Monitoring of the prostate tumour cells redox state and real-time proliferation by novel biophysical techniques and fluorescent staining. \textit{Integrative Biology}. 2012, vol.~4, No~6, p.~672–684. ISSN~1757-9694. Available from: https://dx.doi.org/10.1039/c2ib00157h.

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